Human aortic smooth muscle cell-derived exosomal miR-221/222 inhibits autophagy via a PTEN/Akt signaling pathway in human umbilical vein endothelial cells
Human aortic smooth muscle cell-derived exosomal miR-221/222 inhibits autophagy via a PTEN/Akt signaling pathway in human umbilical vein endothelial cells
复制标题
人主动脉平滑肌细胞来源的外泌体 miR-221/222 通过 PTEN/Akt 信号通路抑制人脐静脉内皮细胞的自噬
DOI：
10.1016/j.bbrc.2016.09.078
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发表时间：
2016-10-14
影响因子：
3.1
通讯作者：
Hu, Rui
中科院分区：
文献类型：
--
作者：
Li, Luocheng;Wang, Zhiwei;Hu, Rui
内皮细胞自噬失调在心血管功能障碍和动脉粥样硬化中起着至关重要的作用。越来越多的证据表明，mirna通过靶向自噬相关基因来调节各种细胞类型的自噬。在本研究中，我们发现人脐静脉内皮细胞（HUVECs）与人主动脉平滑肌细胞（HAoSMCs）共培养可抑制HUVECs的自噬活性。此外，我们分离了由HAoSMCs分泌的外泌体，并证实外泌体含有miR-221/222。我们研究了由haosmc衍生的外泌体转移的miR-221/222在huvec中的作用。这些外泌体诱导HUVECs中miR-221/222表达增加，并下调10号染色体上缺失的磷酸酶和紧张素同源物（PTEN）。双荧光素酶报告基因实验显示miR-221/222可以结合PTEN的3'UTR，这意味着PTEN是miR-221/222的直接靶标。miR-221/222过表达可下调PTEN的表达。然后，我们检测了PTEN、LC3、ATG5、SQSTM1/p62、Beclin-1、Akt和phospho-Akt在转染miR-221/222模拟物和抑制剂的HUVECs中的表达。我们的研究结果表明，miR-221/222过表达抑制PTEN的表达，进而激活Akt信号通路，最终下调LC3II、ATG5和Beclin-1的表达，上调SQSTM1/p62的表达。这种现象可以通过转染miR-221/222抑制剂来逆转。这些数据表明，来自haosmc衍生外泌体的miR-221/222通过调节PTEN/Akt信号通路抑制HUVECs的自噬。(C) 2016 Elsevier Inc.版权所有。
Dysregulation of autophagy in endothelial cells plays a vital role in cardiovascular dysfunction and atherosclerosis. Accumulating evidence shows that miRNAs regulate autophagy in various cell types by targeting autophagy-related genes. In the present study, we found that a co-culture of human umbilical vein endothelial cells (HUVECs) with human aortic smooth muscle cells (HAoSMCs) inhibited autophagy activity in HUVECs. Furthermore, we isolated exosomes secreted by HAoSMCs, and confirmed that the exosomes contain miR-221/222. We investigated the role of miR-221/222 transferred by HAoSMC-derived exosomes in HUVECs. These exosomes induced an increase of miR-221/222 expression and a down-regulation of phosphatase and tensin homolog deleted on chromosome ten (PTEN) in HUVECs. Dual luciferase reporter assays revealed that miR-221/222 could bind to the 3'UTR of PTEN, which implied that PTEN was a direct target of miR-221/222. The expression of PTEN could be down-regulated by miR-221/222 over-expression. Then, we detected the expression of PTEN, LC3, ATG5, SQSTM1/p62, Beclin-1, Akt, and phospho-Akt in HUVECs transfected with miR-221/222 mimics and inhibitors. Our results demonstrated that miR-221/222 overexpression inhibited the expression of PTEN and subsequently activated Akt signaling, and eventually down-regulated the expression of LC3II, ATG5 and Beclin-1, and elevated the expression of SQSTM1/p62. This phenomenon can be reversed by the transfection of miR-221/222 inhibitors. These data suggested that miR-221/222 from HAoSMC-derived exosomes inhibited autophagy in HUVECs by modulating PTEN/Akt signaling pathway. (C) 2016 Elsevier Inc. All rights reserved.